The power of light: Impact on the performance of biocontrol agents under minimal nutrient conditions.

Background: The spectral distribution of light (different wavelength) has recently been identified as an important factor in the dynamics and function of leaf-associated microbes. This study investigated the impact of different wavelength on three commercial biocontrol agents (BCA): Bacillus amyloliquefaciens (BA), Pseudomonas chlororaphis (PC), and Streptomyces griseoviridis (SG). Methods: The impact of light exposure on sole carbon source utilization, biofilm formation, and biosurfactant production by the selected BCA was studied using phenotypic microarray (PM) including 190 sole carbon sources (OmniLog®, PM panels 1 and 2). The BCA were exposed to five monochromatic light conditions (420, 460, 530, 630, and 660 nm) and darkness during incubation, at an intensity of 50 µmol m-2 s-1. Results: Light exposure together with specific carbon source increased respiration in all three BCA. Different wavelengths of light influenced sole carbon utilization for the different BCA, with BA and PC showing increased respiration when exposed to wavelengths within the blue spectrum (420 and 460 nm) while respiration of selected carbon sources by SG increased in the presence of red light (630 and 660 nm). Only one carbon source (capric acid) generated biosurfactant production in all three BCA. A combination of specific wavelength of light and sole carbon source increased biofilm formation in all three BCA. BA showed significantly higher biofilm formation when exposed to blue (460 nm) and green (530 nm) light and propagated in D-sucrose, D-fructose, and dulcitol. PC showed higher biofilm formation when exposed to blue light. Biofilm formation by SG increased when exposed to red light (630 nm) and propagated in citraconic acid. Conclusion: To increase attachment and success in BCA introduced into the phyllosphere, a suitable combination of light quality and nutrient conditions could be used.

Odor-mediated response of gravid Aedes aegypti to mosquito-associated symbiotic bacteria.

Complex oviposition decisions allow gravid Aedes aegypti mosquitoes to select suitable sites for egg-laying to increase the probability that their progeny will thrive. The bacterial communities present in larval niches influence mosquito oviposition behavior, and gravid mosquitoes transmit key microbial associates to breeding sites during oviposition. Our study evaluated whether symbiotic Klebsiella sp., which are strongly associated with mosquitoes, emit volatiles that affect mosquito oviposition decisions. Dual-choice behavioral assays demonstrated that volatile organic compounds emitted by Klebsiella sp. induce a preference in oviposition decisions by Ae. aegypti. Bacterial headspace volatiles were sampled by solid-phase microextraction, and subsequent combined gas chromatography and electroantennogram detection analysis, revealed that the antennae of gravid females detect two compounds present in the Klebsiella sp. headspace. These compounds were identified by gas chromatography and mass spectrometry as 2-ethyl hexanol and 2,4-di­tert-butylphenol. The binary blend of these compounds elicited a dose-dependent egg-laying preference by gravid mosquitoes. We propose that bacterial symbionts, which are associated with gravid mosquitoes and may be transferred to aquatic habitats during egg-laying, together with their volatiles act as oviposition cues indicating the suitability of active breeding sites to conspecific females.

Seed inoculation with antagonistic bacteria limits occurrence of E. coli O157:H7gfp + on baby spinach leaves.

BACKROUND: During the last decades, outbreaks of foodborne illnesses have increasingly been linked to fresh and/or minimally processed fruit and vegetables. Enterohemorrhagic Escherichia coli was the causal agent for major outbreaks in Europe with leafy green vegetables and sprouts. To improve food safety, microbial antagonism has received attention during recent years and could be one of the solution to prevent contamination of food borne pathogens on fresh produce. Here we investigate the antagonistic effect of three bacterial strains (Pseudomonas orientalis, P. flavescens and Rhodococcus sp.) isolated from spinach leaves against E. coli O157:H7gfp + under laboratory and greenhouse conditions. RESULTS: Our results shows that significantly less culturable E.coli O157:H7gfp + were retrieved from the spinach canopy subjected to antagonist seed treatment than canopy inoculation. Seeds inoculated with Rhodococcus sp. significantly reduced growth of E. coli O157:H7gfp + compared with the other antagonists. The result from the in vitro study shows a significant reduction of growth of E. coli O157:H7gfp+, but only after 44 h when E. coli O157:H7gfp + was propagated in a mixture of spent media from all three antagonists. CONCLUSIONS: The antagonistic effect on phyllospheric E.coli O157:H7gfp + observed after seed inoculation with Rhodococcus sp. might be an indication of induced resistance mechanism in the crop. In addition, there was a small reduction of culturable E.coli O157:H7gfp + when propagated in spent media from all three antagonists. Nutritional conditions rather than metabolites formed by the three chosen organisms appear to be critical for controlling E. coli O157:H7gfp+.

Comparative immunomodulatory effects in mice and in human dendritic cells of five bacterial strains selected for biocontrol of leafy green vegetables.

The market for ready-to eat vegetables is increasing, but unfortunately so do the numbers of food-borne illness outbreaks related to these products. A previous study has identified bacterial strains suitable for biocontrol of leafy green vegetables to reduce the exposure to pathogens in these products. As a tentative safety evaluation, five selected strains (Rhodococcus cerastii MR5x, Bacillus coagulans LMG P-32205, Bacillus coagulans LMG P-32206, Pseudomonas cedrina LMG P-32207 and Pseudomonas punonensis LMG P-32204) were individually compared for immunomodulating effects in mice and in human monocyte-derived dendritic cells (MoDCs). Mice receiving the two B. coagulans strains consistently resemble the immunological response of the normal control, and no, or low, cell activation and pro-inflammatory cytokine expression was observed in MoDCs exposed to B. coagulans strains. However, different responses were seen in the two models for the Gram-negative P. cedrina and the Gram-positive R. cerastii. Moreover, P. punonensis and B. coagulans increased the microbiota diversity in mice as seen by the Shannon-Wiener index. In conclusion, the two strains of B. coagulans showed an immunological response that indicate that they lack pathogenic abilities, thus encouraging further safety evaluation and showing great potential to be used as biocontrol agents on leafy green vegetables.

Blue Light Inhibits E. coli, but Decisive Parameters Remain Hidden in the Dark: Systematic Review and Meta-Analysis.

Blue light (400-500 nm) alleviates overexposure risks associated to UV light and has therefore gained increased interest in multiple applications. This meta-analysis deals with decontamination of E. coli through the use of blue light based from nine recent publications identified via a systematic literature search. In these studies, various pathogenic and non-pathogenic E. coli strains grown in nutritional broths were exposed to wavelengths ranging from 395 to 460 nm. Five meta-analyses were performed using Cochrane's software for meta-analyses (Review Manager): one including all studies to estimate the effect of E. coli reduction and four subgroup-analyses considering reported intensities, wavelengths, exposure dose as well as serovars/pathovars. Random effects models were used. All included studies used colony-forming units to estimate the impact of E. coli reduction. None of the included studies involved an organic matrix (e.g., skin, food related surface). Exposure to blue light had a significant and large reducing effect on viable counts of E. coli. However, substantial heterogeneity across studies was observed. Among subgroups, reported intensity and wavelength showed the clearest impact on E. coli reduction. With respect to the reported exposure dose, the picture across the spectrum was scattered, but effect sizes tend to increase with increasing exposure dose. Substantial heterogeneity was also present with respect to all serovar/pathovar subgroups among the included studies. The present body of reports does not display a strong basis for recommendation of relevant intensities, wavelengths and exposure doses for superficial blue light decontamination in medical or food safety contexts. A serious shortcoming in most studies is the absence of a clear documentation of inoculum preparation and of study parameters. We suggest improvement for study protocols for future investigations.

Dining in Blue Light Impairs the Appetite of Some Leaf Epiphytes.

Background: The phyllosphere is subjected to fluctuating abiotic conditions. This study examined the phenotypic plasticity (PP) of four selected non-phototrophic phyllosphere bacteria [control strain: Pseudomonas sp. DR 5-09; Pseudomonas agarici, Bacillus thuringiensis serovar israeliensis (Bti), and Streptomyces griseoviridis (SG)] regarding their respiration patterns and surfactant activity as affected by light spectrum and nutrient supply. Methods: The PP of the strains was examined under four light regimes [darkness (control); monochromatic light-emitting diodes (LED) at 460 nm (blue) and 660 nm (red); continuously polychromatic white LEDs], in the presence of 379 substrates and conditions. Results: Light treatment affected the studied bacterial strains regarding substrate utilization (Pseudomonas strains > SG > Bti). Blue LEDs provoked the most pronounced impact on the phenotypic reaction norms of the Pseudomonas strains and Bti. The two Gram-positive strains Bti and SG, respectively, revealed inconsistent biosurfactant formation in all cases. Biosurfactant formation by both Pseudomonas strains was supported by most substrates incubated in darkness, and blue LED exposure altered the surface activity profoundly. Blue and white LEDs enhanced biofilm formation in PA in highly utilized C-sources. Putative blue light receptor proteins were found in both Pseudomonas strains, showing 91% similarity with the sequence from NCBI accession number WP_064119393. Conclusion: Light quality-nutrient interactions affect biosurfactant activity and biofilm formation of some non-phototrophic phyllosphere bacteria and are, thus, crucial for dynamics of the phyllosphere microbiome.

Immune response and gut microbiota of mice on a diet mimicking eating habits of elderly with risk of malnutrition development.

The number of elderlies is increasing but prevalence of malnutrition has been reported. The aim of the study was to determine the significance of short-term nutritional deficiencies in mice. Immune status was assessed through flow cytometry of leucocytes in Peyer's patches (PP) and mesenteric lymph nodes (MLN), and intestinal microbiota was evaluated by terminal restriction fragment length polymorphism (T-RFLP). C57BL/6NCrl mice fed standard diet (StD) or experimental diet high in fat, and low in carbohydrates, protein, fibre, vitamins, and minerals (ExpD) for 2 or 4 weeks. ExpD-animals gained less weight, increased liver lipids, and developed splenomegaly. Diet affected regulatory T-cells, gut homing receptors and TLR2 and TLR4 in PP and MLN and the microbiota was influenced. Partial least squares models on flow cytometry- and T-RFLP data demonstrated correlations between microbial communities and immune phenotyping. Our model shows similarities to malnourished elderly and interactions between intestinal bacteria and the immune system.

Commercial wash of leafy vegetables do not significantly decrease bacterial load but leads to shifts in bacterial species composition.

Production of leafy vegetables for the "Ready-to-eat"-market has vastly increased the last 20 years, and consumption of these minimally processed vegetables has led to outbreaks of food-borne diseases. Contamination of leafy vegetables can occur throughout the production chain, and therefore washing of the produce has become a standard in commercial processing. This study explores the bacterial communities of spinach (Spinacia oleracea) and rocket (Diplotaxis tenuifolia) in a commercial setting in order to identify potential contamination events, and to investigate effects on bacterial load by commercial processing. Samples were taken in field, after washing of the produce and at the end of shelf-life. This study found that the bacterial community composition and diversity changed significantly from the first harvest to the end of shelf-life, where the core microbiome from the first to the last sampling constituted <2% of all OTUs. While washing of the produce had no reducing effect on bacterial load compared to unwashed, washing led to a change in species composition. As the leaves entered the cold chain after harvest, a rise was seen in the relative abundance of spoilage bacteria. E. coli was detected after the washing indicating issues of cross-contamination in the wash water.

High-throughput method for detection and quantification of lesions on leaf scale based on trypan blue staining and digital image analysis.

BACKGROUND: Field-grown leafy vegetables can be damaged by biotic and abiotic factors, or mechanically damaged by farming practices. Available methods to evaluate leaf tissue damage mainly rely on colour differentiation between healthy and damaged tissues. Alternatively, sophisticated equipment such as microscopy and hyperspectral cameras can be employed. Depending on the causal factor, colour change in the wounded area is not always induced and, by the time symptoms become visible, a plant can already be severely affected. To accurately detect and quantify damage on leaf scale, including microlesions, reliable differentiation between healthy and damaged tissue is essential. We stained whole leaves with trypan blue dye, which traverses compromised cell membranes but is not absorbed in viable cells, followed by automated quantification of damage on leaf scale. RESULTS: We present a robust, fast and sensitive method for leaf-scale visualisation, accurate automated extraction and measurement of damaged area on leaves of leafy vegetables. The image analysis pipeline we developed automatically identifies leaf area and individual stained (lesion) areas down to cell level. As proof of principle, we tested the methodology for damage detection and quantification on two field-grown leafy vegetable species, spinach and Swiss chard. CONCLUSIONS: Our novel lesion quantification method can be used for detection of large (macro) or single-cell (micro) lesions on leaf scale, enabling quantification of lesions at any stage and without requiring symptoms to be in the visible spectrum. Quantifying the wounded area on leaf scale is necessary for generating prediction models for economic losses and produce shelf-life. In addition, risk assessments are based on accurate prediction of the relationship between leaf damage and infection rates by opportunistic pathogens and our method helps determine the severity of leaf damage at fine resolution.

Social Stratification, Diet Diversity and Malnutrition among Preschoolers: A Survey of Addis Ababa, Ethiopia.

In Sub-Saharan Africa, being overweight in childhood is rapidly rising while stunting is still remaining at unacceptable levels. A key contributor to this double burden of malnutrition is dietary changes associated with nutrition transition. Although the importance of socio-economic drivers is known, there is limited knowledge about their stratification and relative importance to diet and to different forms of malnutrition. The aim of this study was to assess diet diversity and malnutrition in preschoolers and evaluate the relative importance of socioeconomic resources. Households with children under five (5467) were enrolled using a multi-stage sampling procedure. Standardized tools and procedures were used to collect data on diet, anthropometry and socio-economic factors. Multivariable analysis with cluster adjustment was performed. The prevalence of stunting was 19.6% (18.5-20.6), wasting 3.2% (2.8-3.7), and overweight/obesity 11.4% (10.6-12.2). Stunting, overweight, wasting and limited diet diversity was present in all social strata. Low maternal education was associated with an increased risk of stunting (Adjusted odds ratio (AOR): 1.8; 1.4-2.2), limited diet diversity (AOR: 0.33; 0.26-0.42) and reduced odds of being overweight (AOR: 0.61; 0.44-0.84). Preschoolers in Addis Ababa have limited quality diets and suffer from both under- and over-nutrition. Maternal education was an important explanatory factor for stunting and being overweight. Interventions that promote diet quality for the undernourished whilst also addressing the burgeoning problem of being overweight are needed.

Size Matters: Biological and Food Safety Relevance of Leaf Damage for Colonization of Escherichia coli O157:H7 gfp.

This study examined the biological and food safety relevance of leaf lesions for potential invasion of food pathogens into the plant tissue (internalization). This was done by determining the role of artificial leaf damage in terms of damaged leaf area on proliferation of E. coli O157:H7 gfp+. In a two-factorial experiment, unwashed fresh baby leaf spinach (Spinacia oleracea L.) was subjected to four damage levels (undamaged, low, moderate, high damage; factor 1) and three incubation intervals (0, 1, 2 days post-inoculation; factor 2). Individual leaves were immersed for 15 s in a suspension loaded with E. coli O157:H7 gfp+ (106 CFU × mL-1). The leaves were analyzed individually using image analysis tools to quantify leaf area and number and size of lesions, and using confocal laser scanning and scanning electron microscopy to visualize leaf lesions and presence of the introduced E. coli strain on and within the leaf tissue. Prevalence of E. coli O157:H7 gfp+ was assessed using a culture-dependent technique. The results showed that size of individual lesions and damaged leaf area affected depth of invasion into plant tissue, dispersal to adjacent areas, and number of culturable E. coli O157:H7 gfp+ directly after inoculation. Differences in numbers of the inoculant retrieved from leaf macerate evened out from 2 days post-inoculation, indicating rapid proliferation during the first day post-inoculation. Leaf weight was a crucial factor, as lighter spinach leaves (most likely younger leaves) were more prone to harbor E. coli O157:H7 gfp+, irrespective of damage level. At the high inoculum density used, the risk of consumers' infection was almost 100%, irrespective of incubation duration or damage level. Even macroscopically intact leaves showed a high risk for infection. These results suggest that the risk to consumers is correlated with how early in the food chain the leaves are contaminated, and the degree of leaf damage. These findings should be taken into account in different steps of leafy green processing. Further attention should be paid to the fate of viable, but non-culturable, shiga-toxigenic E. coli on and in ready-to-eat leafy vegetables.

Leaf mineral content govern microbial community structure in the phyllosphere of spinach (Spinacia oleracea) and rocket (Diplotaxis tenuifolia).

The plant microbiome is an important factor for plant health and productivity. While the impact of nitrogen (N) availability for plant growth and development is well established, its influence on the microbial phyllosphere community structure is unknown. We hypothesize that nitrogen impacts the growth and abundance of several microorganisms on the leaf surface. The bacterial and fungal communities of baby leaf spinach (Spinacia oleracea), and rocket (Diplotaxis tenuifolia) were investigated in a field trial for two years in a commercial setting. Nitrogen fertilizer was tested in four doses (basic nitrogen, basic + suboptimal, basic + commercial, basic + excess) with six replicates in each. Culture-independent (Illumina sequencing) and culture-dependent (viable count and identification of bacterial isolates) community studies were combined with monitoring of plant physiology and site weather conditions. This study found that alpha diversity of bacterial communities decreased in response to increasing nitrogen fertilizer dose, whereas viable counts showed no differences. Correspondingly, fungal communities of the spinach phyllosphere showed a decreasing pattern, whereas the decreasing diversity of fungal communities of rocket was not significant. Plant species and effects of annual variations on microbiome structure were observed for bacterial and fungal communities on both spinach and rocket. This study provides novel insights on the impact of nitrogen fertilizer regime on a nutrient scarce habitat, the phyllosphere.

Season and Species: Two Possible Hurdles for Reducing the Food Safety Risk of Escherichia coli O157 Contamination of Leafy Vegetables.

The food safety risk of Shiga toxin-producing Escherichia coli (STEC) infection per serving of leafy vegetables was investigated using a quantitative microbial risk assessment (QMRA) approach. The estimated level of E. coli O157 contamination was based on observed numbers of Enterobacteriaceae and E. coli on leafy vegetables grown and processed in southern Sweden from 2014 to 2016. Samples were collected before harvest, after washing, and at the end of shelf life. The observed counts were combined with data on the ratio of E. coli to E. coli O157 taken from earlier studies to estimate the probability of illness. The risks of STEC infection associated with species, either spinach ( Spinacia oleracea) or rocket ( Diplotaxis tenuifolia), growing season (spring or autumn), and washing (washed or not washed) were then evaluated. The results indicated that leafy vegetable species and growing season could be possible hurdles for reducing the food safety risk of STEC infection. At harvest, the probability of infection was 87% lower when consuming rocket compared with spinach and 90% lower when consuming leafy vegetables grown in spring compared with autumn. These relative risk reductions remained consistent even with other serving sizes and dose-response models. The lowest risk of STEC infection was associated with leafy vegetables early in the production chain, i.e., before harvest, while the risk increased during storage and processing. Consequently, the highest risk was observed when leafy vegetables were consumed at the end of shelf life. Washing had no effect on the food safety risk of STEC infection in this study. To improve the quality of QMRA, there is a need for additional data on the relationship between indicator organisms that can be easily enumerated (e.g., E. coli and Enterobacteriaceae) and E. coli strains that can cause STEC infection (e.g., E. coli O157) but are difficult to identify in food samples such as leafy vegetables.

Mixed blessings: A qualitative exploration of mothers' experience of child care and feeding in the rapidly urbanizing city of Addis Ababa, Ethiopia.

Many studies have drawn attention to the vital role mothers have in safeguarding the health and nutritional wellbeing of their children. However, little is known about mothers' experiences and the challenges they face in fulfilling this role in rapidly urbanizing cities in Africa. This study aims to explore child care and feeding practices of mothers with children under five years of age in Addis Ababa, Ethiopia. This qualitative study was conducted using a semi-structured interview guide. A total of thirty-six interviews were conducted with purposively selected participants. All interviews were audio recorded, transcribed verbatim and translated for analysis. We used a thematic analysis approach, which was guided by a resilience framework. The findings are presented as three major themes. 1) 'Mixed blessings-balancing motherhood's expectations'. While mothers identified positively with the social recognition and sense of fulfillment of being a 'good mother', they were ambivalent/torn about earning the necessary income from outside work and fulfilling their duties at home. 2) 'Instabilities due to rampant urban sprawl'. While women expressed a keen desire to balance work and motherhood, the disintegrating social capital, due to large in-migration, market fluctuations and abrupt/forced resettlements to new housing units had left mothers without support for childcare, stressed and exhausted. 3) 'Anchored by faith: a source of resilience to cope with adversities'. In the face of the multiple adversities, mothers cited their strong faith as their most reliable foundation for their resilience. In summary, the societal and environmental changes accompanying the rapid urbanization in low income settings makes combining child care and working outside the home very challenging for mothers. As a result they suffer from fatigue and feelings of isolation. Efforts to improve child feeding and care in urban low-income settings need to consider context appropriate strategies that support mothers with small children.

The Hurdle Approach-A Holistic Concept for Controlling Food Safety Risks Associated With Pathogenic Bacterial Contamination of Leafy Green Vegetables. A Review.

Consumers appreciate leafy green vegetables such as baby leaves for their convenience and wholesomeness and for adding a variety of tastes and colors to their plate. In Western cuisine, leafy green vegetables are usually eaten fresh and raw, with no step in the long chain from seed to consumption where potentially harmful microorganisms could be completely eliminated, e.g., through heating. A concerning trend in recent years is disease outbreaks caused by various leafy vegetable crops and one of the most important foodborne pathogens in this context is Shiga toxin-producing Escherichia coli (STEC). Other pathogens such as Salmonella, Shigella, Yersinia enterocolitica and Listeria monocytogenes should also be considered in disease risk analysis, as they have been implicated in outbreaks associated with leafy greens. These pathogens may enter the horticultural value network during primary production in field or greenhouse via irrigation, at harvest, during processing and distribution or in the home kitchen/restaurant. The hurdle approach involves combining several mitigating approaches, each of which is insufficient on its own, to control or even eliminate pathogens in food products. Since the food chain system for leafy green vegetables contains no absolute kill step for pathogens, use of hurdles at critical points could enable control of pathogens that pose a human health risk. Hurdles should be combined so as to decrease the risk due to pathogenic microbes and also to improve microbial stability, shelf-life, nutritional properties and sensory quality of leafy vegetables. The hurdle toolbox includes different options, such as physical, physiochemical and microbial hurdles. The goal for leafy green vegetables is multi-target preservation through intelligently applied hurdles. This review describes hurdles that could be used for leafy green vegetables and their biological basis, and identifies prospective hurdles that need attention in future research.

What Influences Urban Mothers' Decisions on What to Feed Their Children Aged Under Five-The Case of Addis Ababa, Ethiopia.

Mothers carry the prime responsibility for childcare and feeding in low-income countries. Understanding their experiences in providing food for their children is paramount to informing efforts to improve the nutritional status of children. Such information is lacking in Sub-Saharan Africa. To understand what influences urban mothers' food acquisition and their motivations for selecting food for their children, 36 in-depth interviews were carried out with mothers having children under five years of age. Interviews were conducted in the local language, audio-recorded, transcribed, and translated into English. Data were analyzed using thematic analysis which led to the identification of four major themes: mothers give-in to a child-driven diet; quick-fix versus the privilege of planning; keen awareness on food safety, nutrition, and diet diversity; and social, familial, and cultural influences. The findings indicate that child feeding practices are influenced by interlinked social and environmental factors. Hence, nutrition education campaigns should focus on targeting not only families but also their children. Attention should also be given to food safety regulations, as well as to the much-needed support of mothers who are struggling to ensure their children's survival in low-income countries.

Light spectrum modifies the utilization pattern of energy sources in Pseudomonas sp. DR 5-09.

Despite the overruling impact of light in the phyllosphere, little is known regarding the influence of light spectra on non-phototrophic bacteria colonizing the leaf surface. We developed an in vitro method to study phenotypic profile responses of bacterial pure cultures to different bands of the visible light spectrum using monochromatic (blue: 460 nm; red: 660 nm) and polychromatic (white: 350-990 nm) LEDs, by modification and optimization of a protocol for the Phenotype MicroArray™ technique (Biolog Inc., CA, USA). The new protocol revealed high reproducibility of substrate utilization under all conditions tested. Challenging the non-phototrophic bacterium Pseudomonas sp. DR 5-09 with white, blue, and red light demonstrated that all light treatments affected the respiratory profile differently, with blue LED having the most decisive impact on substrate utilization by impairing respiration of 140 substrates. The respiratory activity was decreased on 23 and 42 substrates under red and white LEDs, respectively, while utilization of one, 16, and 20 substrates increased in the presence of red, blue, and white LEDs, respectively. Interestingly, on four substrates contrasting utilization patterns were found when the bacterium was exposed to different light spectra. Although non-phototrophic bacteria do not rely directly on light as an energy source, Pseudomonas sp. DR 5-09 changed its respiratory activity on various substrates differently when exposed to different lights. Thus, ability to sense and distinguish between different wavelengths even within the visible light spectrum must exist, and leads to differential regulation of substrate usage. With these results, we hypothesize that different light spectra might be a hitherto neglected key stimulus for changes in microbial lifestyle and habits of substrate usage by non-phototrophic phyllospheric microbiota, and thus might essentially stratify leaf microbiota composition and diversity.

Effects of household washing on bacterial load and removal of Escherichia coli from lettuce and "ready-to-eat" salads.

Customer demands for fresh salads are increasing, but leafy green vegetables have also been linked to food-borne illness due to pathogens such as Escherichia coli O157:H7. As a safety measure, consumers often wash leafy vegetables in water before consumption. In this study, we analyzed the efficiency of household washing to reduce the bacterial content. Romaine lettuce and ready-to-eat mixed salad were washed several times in flowing water at different rates and by immersing the leaves in water. Lettuce was also inoculated with E. coli before washing. Only washing in a high flow rate (8 L/min) resulted in statistically significant reductions (p < .05), "Total aerobic count" was reduced by 80%, and Enterobacteriaceae count was reduced by 68% after the first rinse. The number of contaminating E. coli was not significantly reduced. The dominating part of the culturable microbiota of the washed lettuce was identified by rRNA 16S sequencing of randomly picked colonies. The majority belonged to Pseudomonadaceae, but isolates from Enterobacteriaceae and Staphylococcaceaceae were also frequently found. This study shows the inefficiency of tap water washing methods available for the consumer when it comes to removal of bacteria from lettuce. Even after washing, the lettuce contained high levels of bacteria that in a high dose and under certain circumstances may constitute a health risk.

Fate of Listeria monocytogenes , Pathogenic Yersinia enterocolitica , and Escherichia coli O157:H7 gfp+ in Ready-to-Eat Salad during Cold Storage: What Is the Risk to Consumers?

In this study, we investigated the fate of Listeria monocytogenes , pathogenic Yersinia enterocolitica , and Escherichia coli O157:H7 gfp+ inoculated in low numbers into ready-to-eat baby spinach and mixed-ingredient salad (baby spinach with chicken meat). Samples were stored at recommended maximum refrigerator temperature (8°C in Sweden) or at an abuse temperature (15°C) for up to 7 days. Mixed-ingredient salad supported considerable growth when stored at 15°C during shelf life (3 days), with populations of L. monocytogenes , pathogenic Y. enterocolitica , and E. coli O157:H7 gfp+ increasing from less than 2.0 log CFU/g on day 0 to 7.0, 4.0, and 5.6 log CFU/g, respectively. However, when mixed-ingredient salad was stored at 8°C during shelf life, only L. monocytogenes increased significantly, reaching 3.0 log CFU/g within 3 days. In plain baby spinach, only pathogenic Y. enterocolitica populations increased significantly during storage for 7 days, and this was exclusively at an abuse temperature (15°C). Thus, mixing ready-to-eat leafy vegetables with chicken meat strongly influenced levels of inoculated strains during storage. To explore the food safety implications of these findings, bacterial numbers were translated into risks of infection by modeling. The risk of listeriosis (measured as probability of infection) was 16 times higher when consuming a mixed-ingredient salad stored at 8°C at the end of shelf life, or 200,000 times higher when stored at 15°C, compared with when consuming it on the day of inoculation. This indicates that efforts should focus on preventing temperature abuse during storage to mitigate the risk of listeriosis. The storage conditions recommended for mixed-ingredient salads in Sweden (maximum 8°C for 3 days) did not prevent growth of L. monocytogenes in baby spinach mixed with chicken meat. Manufacturers preparing these salads should be aware of this, and recommended storage temperature should be revised downwards to reduce the risk of foodborne disease.